p src  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p src
    P Src, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    plck cell signaling 2101  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc plck cell signaling 2101
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    p src  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc p src
    P Src, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    antibodies against p src family tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc antibodies against p src family tyr416
    Antibodies Against P Src Family Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    pab anti phospho src tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc pab anti phospho src tyr416
    Pab Anti Phospho Src Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    phospho src tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src tyr416
    ( A ) Growth curves with FGF2 (10 ng/mL) and PP1 or PP2 (5 µM) added 1 hour before FGF2 (10 4 cells/cm 2 were plated). ( B ) Clonogenic assays of Y1 cells treated with FGF2 and/or PP1 (10 µM) or PP2 (5 µM) (added 30 minutes before FGF2) for 24 hours. Insets on the bottom show one representative clonogenic assay in the indicated conditions. Values are mean ± s.e.m. (n = 3). ( C ) Microphotographs for morphological analysis of Y1 cells under FCS or FGF2 stimulation for 48 hours, in the presence, where indicated, of PP2 (10 µM). ( D ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of four independent experiments. Phospho-Src (phosphorylated at <t>Tyr416)</t> and total Src were analyzed by immunoblot. Hprt was used as a loading control. ( E ) G0/G1-arrested Y1 cells were stimulated with FCS and/or FGF2 for 12 hours. Cells were pulse-labeled for 1 hour with [ 3 H]-thymidine before harvesting, and the amount of incorporated radioactivity was measured. FCS and FGF2 were added at time 0. PP1 or PP2 were added 1 hour before stimulation with FCS and/or FGF2. [ 3 H]-thymidine incorporation results are presented as the mean ± s.e.m. (n = 2). ( F ) Flow cytometry (FC) histograms show DNA content in Y1 cells after 24 and 48 hours of FCS and FGF2 treatment in G0/G1-starved cells. Cells were treated at time 0 and stained for DNA content and BrdU. FC histograms of DNA content (upper panels) and DNA/BrdU scatterplots (lower panels) of Y1 cells with or without PP2 (- PP2 and+PP2, respectively). Cells were stained with propidium iodide to assess DNA content. Biparametric flow cytometry analysis of DNA content and BrdU incorporation was performed on the same samples as described in the text. BrdU-positive cells were quantified by gates in the BrdU/DNA scatterplots. The upper panel shows the quantification of G0/G1, S and G2/M phases based on DNA content. The lower panel shows the quantification of G0/G1, S and G2/M phases based on DNA content versus BrdU labeling. Quantification of cell cycle phases, gated from the 2N to the 4N population only. Approximately 2×10 4 cells were analyzed. ( G ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of two independent experiments. phospho-ERK1/2 (phosphorylated at Thr202/Tyr204) and total ERK 1/2 were analyzed by immunoblot. Hprt was used as a loading control. Scale bar = 50 µm. SFM, serum-free media; FCS, fetal calf serum.
    Phospho Src Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Fibroblast Growth Factor 2 Causes G2/M Cell Cycle Arrest in Ras-Driven Tumor Cells through a Src-Dependent Pathway"

    Article Title: Fibroblast Growth Factor 2 Causes G2/M Cell Cycle Arrest in Ras-Driven Tumor Cells through a Src-Dependent Pathway

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0072582

    ( A ) Growth curves with FGF2 (10 ng/mL) and PP1 or PP2 (5 µM) added 1 hour before FGF2 (10 4 cells/cm 2 were plated). ( B ) Clonogenic assays of Y1 cells treated with FGF2 and/or PP1 (10 µM) or PP2 (5 µM) (added 30 minutes before FGF2) for 24 hours. Insets on the bottom show one representative clonogenic assay in the indicated conditions. Values are mean ± s.e.m. (n = 3). ( C ) Microphotographs for morphological analysis of Y1 cells under FCS or FGF2 stimulation for 48 hours, in the presence, where indicated, of PP2 (10 µM). ( D ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of four independent experiments. Phospho-Src (phosphorylated at Tyr416) and total Src were analyzed by immunoblot. Hprt was used as a loading control. ( E ) G0/G1-arrested Y1 cells were stimulated with FCS and/or FGF2 for 12 hours. Cells were pulse-labeled for 1 hour with [ 3 H]-thymidine before harvesting, and the amount of incorporated radioactivity was measured. FCS and FGF2 were added at time 0. PP1 or PP2 were added 1 hour before stimulation with FCS and/or FGF2. [ 3 H]-thymidine incorporation results are presented as the mean ± s.e.m. (n = 2). ( F ) Flow cytometry (FC) histograms show DNA content in Y1 cells after 24 and 48 hours of FCS and FGF2 treatment in G0/G1-starved cells. Cells were treated at time 0 and stained for DNA content and BrdU. FC histograms of DNA content (upper panels) and DNA/BrdU scatterplots (lower panels) of Y1 cells with or without PP2 (- PP2 and+PP2, respectively). Cells were stained with propidium iodide to assess DNA content. Biparametric flow cytometry analysis of DNA content and BrdU incorporation was performed on the same samples as described in the text. BrdU-positive cells were quantified by gates in the BrdU/DNA scatterplots. The upper panel shows the quantification of G0/G1, S and G2/M phases based on DNA content. The lower panel shows the quantification of G0/G1, S and G2/M phases based on DNA content versus BrdU labeling. Quantification of cell cycle phases, gated from the 2N to the 4N population only. Approximately 2×10 4 cells were analyzed. ( G ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of two independent experiments. phospho-ERK1/2 (phosphorylated at Thr202/Tyr204) and total ERK 1/2 were analyzed by immunoblot. Hprt was used as a loading control. Scale bar = 50 µm. SFM, serum-free media; FCS, fetal calf serum.
    Figure Legend Snippet: ( A ) Growth curves with FGF2 (10 ng/mL) and PP1 or PP2 (5 µM) added 1 hour before FGF2 (10 4 cells/cm 2 were plated). ( B ) Clonogenic assays of Y1 cells treated with FGF2 and/or PP1 (10 µM) or PP2 (5 µM) (added 30 minutes before FGF2) for 24 hours. Insets on the bottom show one representative clonogenic assay in the indicated conditions. Values are mean ± s.e.m. (n = 3). ( C ) Microphotographs for morphological analysis of Y1 cells under FCS or FGF2 stimulation for 48 hours, in the presence, where indicated, of PP2 (10 µM). ( D ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of four independent experiments. Phospho-Src (phosphorylated at Tyr416) and total Src were analyzed by immunoblot. Hprt was used as a loading control. ( E ) G0/G1-arrested Y1 cells were stimulated with FCS and/or FGF2 for 12 hours. Cells were pulse-labeled for 1 hour with [ 3 H]-thymidine before harvesting, and the amount of incorporated radioactivity was measured. FCS and FGF2 were added at time 0. PP1 or PP2 were added 1 hour before stimulation with FCS and/or FGF2. [ 3 H]-thymidine incorporation results are presented as the mean ± s.e.m. (n = 2). ( F ) Flow cytometry (FC) histograms show DNA content in Y1 cells after 24 and 48 hours of FCS and FGF2 treatment in G0/G1-starved cells. Cells were treated at time 0 and stained for DNA content and BrdU. FC histograms of DNA content (upper panels) and DNA/BrdU scatterplots (lower panels) of Y1 cells with or without PP2 (- PP2 and+PP2, respectively). Cells were stained with propidium iodide to assess DNA content. Biparametric flow cytometry analysis of DNA content and BrdU incorporation was performed on the same samples as described in the text. BrdU-positive cells were quantified by gates in the BrdU/DNA scatterplots. The upper panel shows the quantification of G0/G1, S and G2/M phases based on DNA content. The lower panel shows the quantification of G0/G1, S and G2/M phases based on DNA content versus BrdU labeling. Quantification of cell cycle phases, gated from the 2N to the 4N population only. Approximately 2×10 4 cells were analyzed. ( G ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of two independent experiments. phospho-ERK1/2 (phosphorylated at Thr202/Tyr204) and total ERK 1/2 were analyzed by immunoblot. Hprt was used as a loading control. Scale bar = 50 µm. SFM, serum-free media; FCS, fetal calf serum.

    Techniques Used: Clonogenic Assay, Western Blot, Labeling, Radioactivity, Flow Cytometry, Staining, BrdU Incorporation Assay

    anti phospho src family tyr 416 antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho src family tyr 416 antibody
    Anti Phospho Src Family Tyr 416 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti src antibody 32g6  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti src antibody 32g6
    Anti Src Antibody 32g6, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    anti phospho src family ptyr416 rabbit polyclonal antibody  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc anti phospho src family ptyr416 rabbit polyclonal antibody
    The S609A mutation inhibits activation of Src family kinases. ( a ) 1b9 or S609A cells were stimulated by ristocetin in the presence or absence of VWF, then solubilized and subjected to Western blot analysis with anti-Src and anti-phospho-Src family <t>(pTyr416)</t> antibody, respectively. The immunoblot is representative of 3 different experiments; ( b ) Quantitative data from 3 different experiments (mean ± SD) are demonstrated. The relative activation index of Src equals arbitrary quantitation of phospho-Src (pTyr416)/arbitrary quantitation of total Src.
    Anti Phospho Src Family Ptyr416 Rabbit Polyclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Role of 14-3-3ζ in Platelet Glycoprotein Ibα-von Willebrand Factor Interaction-Induced Signaling"

    Article Title: Role of 14-3-3ζ in Platelet Glycoprotein Ibα-von Willebrand Factor Interaction-Induced Signaling

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms13055364

    The S609A mutation inhibits activation of Src family kinases. ( a ) 1b9 or S609A cells were stimulated by ristocetin in the presence or absence of VWF, then solubilized and subjected to Western blot analysis with anti-Src and anti-phospho-Src family (pTyr416) antibody, respectively. The immunoblot is representative of 3 different experiments; ( b ) Quantitative data from 3 different experiments (mean ± SD) are demonstrated. The relative activation index of Src equals arbitrary quantitation of phospho-Src (pTyr416)/arbitrary quantitation of total Src.
    Figure Legend Snippet: The S609A mutation inhibits activation of Src family kinases. ( a ) 1b9 or S609A cells were stimulated by ristocetin in the presence or absence of VWF, then solubilized and subjected to Western blot analysis with anti-Src and anti-phospho-Src family (pTyr416) antibody, respectively. The immunoblot is representative of 3 different experiments; ( b ) Quantitative data from 3 different experiments (mean ± SD) are demonstrated. The relative activation index of Src equals arbitrary quantitation of phospho-Src (pTyr416)/arbitrary quantitation of total Src.

    Techniques Used: Mutagenesis, Activation Assay, Western Blot, Quantitation Assay

    phospho src  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src
    Animals were treated with 0.5% DMSO (control) or PD98059 at 300 µg/kg. Immunoblots and the respective densitometries are shown <t>for</t> <t>p-ERK1/2,</t> ERK1, <t>p-Src</t> and Src. Thirty µg of total protein were applied to each lane and each band is representative of one animal. β-actin was used as loading control. Relative densitometries as ratio of p-ERK/ERK1 and p-Src/Src are shown as fold change of control group and are presented by bars as means±SD (n = 3/group). *p<0.05 when comparing PD98059-treated group to control. Assays were conducted in duplicates.
    Phospho Src, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "MAPK Signaling Pathway Regulates p27 Phosphorylation at Threonin 187 as Part of the Mechanism Triggered by Early-Weaning to Induce Cell Proliferation in Rat Gastric Mucosa"

    Article Title: MAPK Signaling Pathway Regulates p27 Phosphorylation at Threonin 187 as Part of the Mechanism Triggered by Early-Weaning to Induce Cell Proliferation in Rat Gastric Mucosa

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0066651

    Animals were treated with 0.5% DMSO (control) or PD98059 at 300 µg/kg. Immunoblots and the respective densitometries are shown for p-ERK1/2, ERK1, p-Src and Src. Thirty µg of total protein were applied to each lane and each band is representative of one animal. β-actin was used as loading control. Relative densitometries as ratio of p-ERK/ERK1 and p-Src/Src are shown as fold change of control group and are presented by bars as means±SD (n = 3/group). *p<0.05 when comparing PD98059-treated group to control. Assays were conducted in duplicates.
    Figure Legend Snippet: Animals were treated with 0.5% DMSO (control) or PD98059 at 300 µg/kg. Immunoblots and the respective densitometries are shown for p-ERK1/2, ERK1, p-Src and Src. Thirty µg of total protein were applied to each lane and each band is representative of one animal. β-actin was used as loading control. Relative densitometries as ratio of p-ERK/ERK1 and p-Src/Src are shown as fold change of control group and are presented by bars as means±SD (n = 3/group). *p<0.05 when comparing PD98059-treated group to control. Assays were conducted in duplicates.

    Techniques Used: Western Blot

    phospho src tyr416  (Cell Signaling Technology Inc)


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    Cell Signaling Technology Inc phospho src tyr416
    Phospho Src Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho src tyr416
    ( A ) Growth curves with FGF2 (10 ng/mL) and PP1 or PP2 (5 µM) added 1 hour before FGF2 (10 4 cells/cm 2 were plated). ( B ) Clonogenic assays of Y1 cells treated with FGF2 and/or PP1 (10 µM) or PP2 (5 µM) (added 30 minutes before FGF2) for 24 hours. Insets on the bottom show one representative clonogenic assay in the indicated conditions. Values are mean ± s.e.m. (n = 3). ( C ) Microphotographs for morphological analysis of Y1 cells under FCS or FGF2 stimulation for 48 hours, in the presence, where indicated, of PP2 (10 µM). ( D ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of four independent experiments. Phospho-Src (phosphorylated at <t>Tyr416)</t> and total Src were analyzed by immunoblot. Hprt was used as a loading control. ( E ) G0/G1-arrested Y1 cells were stimulated with FCS and/or FGF2 for 12 hours. Cells were pulse-labeled for 1 hour with [ 3 H]-thymidine before harvesting, and the amount of incorporated radioactivity was measured. FCS and FGF2 were added at time 0. PP1 or PP2 were added 1 hour before stimulation with FCS and/or FGF2. [ 3 H]-thymidine incorporation results are presented as the mean ± s.e.m. (n = 2). ( F ) Flow cytometry (FC) histograms show DNA content in Y1 cells after 24 and 48 hours of FCS and FGF2 treatment in G0/G1-starved cells. Cells were treated at time 0 and stained for DNA content and BrdU. FC histograms of DNA content (upper panels) and DNA/BrdU scatterplots (lower panels) of Y1 cells with or without PP2 (- PP2 and+PP2, respectively). Cells were stained with propidium iodide to assess DNA content. Biparametric flow cytometry analysis of DNA content and BrdU incorporation was performed on the same samples as described in the text. BrdU-positive cells were quantified by gates in the BrdU/DNA scatterplots. The upper panel shows the quantification of G0/G1, S and G2/M phases based on DNA content. The lower panel shows the quantification of G0/G1, S and G2/M phases based on DNA content versus BrdU labeling. Quantification of cell cycle phases, gated from the 2N to the 4N population only. Approximately 2×10 4 cells were analyzed. ( G ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of two independent experiments. phospho-ERK1/2 (phosphorylated at Thr202/Tyr204) and total ERK 1/2 were analyzed by immunoblot. Hprt was used as a loading control. Scale bar = 50 µm. SFM, serum-free media; FCS, fetal calf serum.
    Phospho Src Tyr416, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Cell Signaling Technology Inc anti phospho src family tyr 416 antibody
    ( A ) Growth curves with FGF2 (10 ng/mL) and PP1 or PP2 (5 µM) added 1 hour before FGF2 (10 4 cells/cm 2 were plated). ( B ) Clonogenic assays of Y1 cells treated with FGF2 and/or PP1 (10 µM) or PP2 (5 µM) (added 30 minutes before FGF2) for 24 hours. Insets on the bottom show one representative clonogenic assay in the indicated conditions. Values are mean ± s.e.m. (n = 3). ( C ) Microphotographs for morphological analysis of Y1 cells under FCS or FGF2 stimulation for 48 hours, in the presence, where indicated, of PP2 (10 µM). ( D ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of four independent experiments. Phospho-Src (phosphorylated at <t>Tyr416)</t> and total Src were analyzed by immunoblot. Hprt was used as a loading control. ( E ) G0/G1-arrested Y1 cells were stimulated with FCS and/or FGF2 for 12 hours. Cells were pulse-labeled for 1 hour with [ 3 H]-thymidine before harvesting, and the amount of incorporated radioactivity was measured. FCS and FGF2 were added at time 0. PP1 or PP2 were added 1 hour before stimulation with FCS and/or FGF2. [ 3 H]-thymidine incorporation results are presented as the mean ± s.e.m. (n = 2). ( F ) Flow cytometry (FC) histograms show DNA content in Y1 cells after 24 and 48 hours of FCS and FGF2 treatment in G0/G1-starved cells. Cells were treated at time 0 and stained for DNA content and BrdU. FC histograms of DNA content (upper panels) and DNA/BrdU scatterplots (lower panels) of Y1 cells with or without PP2 (- PP2 and+PP2, respectively). Cells were stained with propidium iodide to assess DNA content. Biparametric flow cytometry analysis of DNA content and BrdU incorporation was performed on the same samples as described in the text. BrdU-positive cells were quantified by gates in the BrdU/DNA scatterplots. The upper panel shows the quantification of G0/G1, S and G2/M phases based on DNA content. The lower panel shows the quantification of G0/G1, S and G2/M phases based on DNA content versus BrdU labeling. Quantification of cell cycle phases, gated from the 2N to the 4N population only. Approximately 2×10 4 cells were analyzed. ( G ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of two independent experiments. phospho-ERK1/2 (phosphorylated at Thr202/Tyr204) and total ERK 1/2 were analyzed by immunoblot. Hprt was used as a loading control. Scale bar = 50 µm. SFM, serum-free media; FCS, fetal calf serum.
    Anti Phospho Src Family Tyr 416 Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc anti src antibody 32g6
    ( A ) Growth curves with FGF2 (10 ng/mL) and PP1 or PP2 (5 µM) added 1 hour before FGF2 (10 4 cells/cm 2 were plated). ( B ) Clonogenic assays of Y1 cells treated with FGF2 and/or PP1 (10 µM) or PP2 (5 µM) (added 30 minutes before FGF2) for 24 hours. Insets on the bottom show one representative clonogenic assay in the indicated conditions. Values are mean ± s.e.m. (n = 3). ( C ) Microphotographs for morphological analysis of Y1 cells under FCS or FGF2 stimulation for 48 hours, in the presence, where indicated, of PP2 (10 µM). ( D ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of four independent experiments. Phospho-Src (phosphorylated at <t>Tyr416)</t> and total Src were analyzed by immunoblot. Hprt was used as a loading control. ( E ) G0/G1-arrested Y1 cells were stimulated with FCS and/or FGF2 for 12 hours. Cells were pulse-labeled for 1 hour with [ 3 H]-thymidine before harvesting, and the amount of incorporated radioactivity was measured. FCS and FGF2 were added at time 0. PP1 or PP2 were added 1 hour before stimulation with FCS and/or FGF2. [ 3 H]-thymidine incorporation results are presented as the mean ± s.e.m. (n = 2). ( F ) Flow cytometry (FC) histograms show DNA content in Y1 cells after 24 and 48 hours of FCS and FGF2 treatment in G0/G1-starved cells. Cells were treated at time 0 and stained for DNA content and BrdU. FC histograms of DNA content (upper panels) and DNA/BrdU scatterplots (lower panels) of Y1 cells with or without PP2 (- PP2 and+PP2, respectively). Cells were stained with propidium iodide to assess DNA content. Biparametric flow cytometry analysis of DNA content and BrdU incorporation was performed on the same samples as described in the text. BrdU-positive cells were quantified by gates in the BrdU/DNA scatterplots. The upper panel shows the quantification of G0/G1, S and G2/M phases based on DNA content. The lower panel shows the quantification of G0/G1, S and G2/M phases based on DNA content versus BrdU labeling. Quantification of cell cycle phases, gated from the 2N to the 4N population only. Approximately 2×10 4 cells were analyzed. ( G ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of two independent experiments. phospho-ERK1/2 (phosphorylated at Thr202/Tyr204) and total ERK 1/2 were analyzed by immunoblot. Hprt was used as a loading control. Scale bar = 50 µm. SFM, serum-free media; FCS, fetal calf serum.
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    Cell Signaling Technology Inc anti phospho src family ptyr416 rabbit polyclonal antibody
    The S609A mutation inhibits activation of Src family kinases. ( a ) 1b9 or S609A cells were stimulated by ristocetin in the presence or absence of VWF, then solubilized and subjected to Western blot analysis with anti-Src and anti-phospho-Src family <t>(pTyr416)</t> antibody, respectively. The immunoblot is representative of 3 different experiments; ( b ) Quantitative data from 3 different experiments (mean ± SD) are demonstrated. The relative activation index of Src equals arbitrary quantitation of phospho-Src (pTyr416)/arbitrary quantitation of total Src.
    Anti Phospho Src Family Ptyr416 Rabbit Polyclonal Antibody, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Cell Signaling Technology Inc phospho src
    Animals were treated with 0.5% DMSO (control) or PD98059 at 300 µg/kg. Immunoblots and the respective densitometries are shown <t>for</t> <t>p-ERK1/2,</t> ERK1, <t>p-Src</t> and Src. Thirty µg of total protein were applied to each lane and each band is representative of one animal. β-actin was used as loading control. Relative densitometries as ratio of p-ERK/ERK1 and p-Src/Src are shown as fold change of control group and are presented by bars as means±SD (n = 3/group). *p<0.05 when comparing PD98059-treated group to control. Assays were conducted in duplicates.
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    Image Search Results


    ( A ) Growth curves with FGF2 (10 ng/mL) and PP1 or PP2 (5 µM) added 1 hour before FGF2 (10 4 cells/cm 2 were plated). ( B ) Clonogenic assays of Y1 cells treated with FGF2 and/or PP1 (10 µM) or PP2 (5 µM) (added 30 minutes before FGF2) for 24 hours. Insets on the bottom show one representative clonogenic assay in the indicated conditions. Values are mean ± s.e.m. (n = 3). ( C ) Microphotographs for morphological analysis of Y1 cells under FCS or FGF2 stimulation for 48 hours, in the presence, where indicated, of PP2 (10 µM). ( D ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of four independent experiments. Phospho-Src (phosphorylated at Tyr416) and total Src were analyzed by immunoblot. Hprt was used as a loading control. ( E ) G0/G1-arrested Y1 cells were stimulated with FCS and/or FGF2 for 12 hours. Cells were pulse-labeled for 1 hour with [ 3 H]-thymidine before harvesting, and the amount of incorporated radioactivity was measured. FCS and FGF2 were added at time 0. PP1 or PP2 were added 1 hour before stimulation with FCS and/or FGF2. [ 3 H]-thymidine incorporation results are presented as the mean ± s.e.m. (n = 2). ( F ) Flow cytometry (FC) histograms show DNA content in Y1 cells after 24 and 48 hours of FCS and FGF2 treatment in G0/G1-starved cells. Cells were treated at time 0 and stained for DNA content and BrdU. FC histograms of DNA content (upper panels) and DNA/BrdU scatterplots (lower panels) of Y1 cells with or without PP2 (- PP2 and+PP2, respectively). Cells were stained with propidium iodide to assess DNA content. Biparametric flow cytometry analysis of DNA content and BrdU incorporation was performed on the same samples as described in the text. BrdU-positive cells were quantified by gates in the BrdU/DNA scatterplots. The upper panel shows the quantification of G0/G1, S and G2/M phases based on DNA content. The lower panel shows the quantification of G0/G1, S and G2/M phases based on DNA content versus BrdU labeling. Quantification of cell cycle phases, gated from the 2N to the 4N population only. Approximately 2×10 4 cells were analyzed. ( G ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of two independent experiments. phospho-ERK1/2 (phosphorylated at Thr202/Tyr204) and total ERK 1/2 were analyzed by immunoblot. Hprt was used as a loading control. Scale bar = 50 µm. SFM, serum-free media; FCS, fetal calf serum.

    Journal: PLoS ONE

    Article Title: Fibroblast Growth Factor 2 Causes G2/M Cell Cycle Arrest in Ras-Driven Tumor Cells through a Src-Dependent Pathway

    doi: 10.1371/journal.pone.0072582

    Figure Lengend Snippet: ( A ) Growth curves with FGF2 (10 ng/mL) and PP1 or PP2 (5 µM) added 1 hour before FGF2 (10 4 cells/cm 2 were plated). ( B ) Clonogenic assays of Y1 cells treated with FGF2 and/or PP1 (10 µM) or PP2 (5 µM) (added 30 minutes before FGF2) for 24 hours. Insets on the bottom show one representative clonogenic assay in the indicated conditions. Values are mean ± s.e.m. (n = 3). ( C ) Microphotographs for morphological analysis of Y1 cells under FCS or FGF2 stimulation for 48 hours, in the presence, where indicated, of PP2 (10 µM). ( D ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of four independent experiments. Phospho-Src (phosphorylated at Tyr416) and total Src were analyzed by immunoblot. Hprt was used as a loading control. ( E ) G0/G1-arrested Y1 cells were stimulated with FCS and/or FGF2 for 12 hours. Cells were pulse-labeled for 1 hour with [ 3 H]-thymidine before harvesting, and the amount of incorporated radioactivity was measured. FCS and FGF2 were added at time 0. PP1 or PP2 were added 1 hour before stimulation with FCS and/or FGF2. [ 3 H]-thymidine incorporation results are presented as the mean ± s.e.m. (n = 2). ( F ) Flow cytometry (FC) histograms show DNA content in Y1 cells after 24 and 48 hours of FCS and FGF2 treatment in G0/G1-starved cells. Cells were treated at time 0 and stained for DNA content and BrdU. FC histograms of DNA content (upper panels) and DNA/BrdU scatterplots (lower panels) of Y1 cells with or without PP2 (- PP2 and+PP2, respectively). Cells were stained with propidium iodide to assess DNA content. Biparametric flow cytometry analysis of DNA content and BrdU incorporation was performed on the same samples as described in the text. BrdU-positive cells were quantified by gates in the BrdU/DNA scatterplots. The upper panel shows the quantification of G0/G1, S and G2/M phases based on DNA content. The lower panel shows the quantification of G0/G1, S and G2/M phases based on DNA content versus BrdU labeling. Quantification of cell cycle phases, gated from the 2N to the 4N population only. Approximately 2×10 4 cells were analyzed. ( G ) Y1 cells were synchronized in G0/G1 by serum starvation and stimulated with FCS (10%) and/or FGF2 (10 ng/mL) for 30 minutes in the presence of PP1 or PP2 (10 µM). Data are representative of two independent experiments. phospho-ERK1/2 (phosphorylated at Thr202/Tyr204) and total ERK 1/2 were analyzed by immunoblot. Hprt was used as a loading control. Scale bar = 50 µm. SFM, serum-free media; FCS, fetal calf serum.

    Article Snippet: Membranes were blocked for 1 hour in TBS-T buffer (150 mM sodium chloride, 50 mM Tris [pH 8], and 0.1% Tween 20) containing 5% nonfat milk and were then incubated with antibodies recognizing AKT (#9272), phospho-AKT Ser473 (#9272), ERK1/2 (#9102), phospho-ERK1/2 Thr202/Tyr204 (#9101L), phospho-Src Tyr416 (#2101), total Src (#2110) (all from Cell Signaling), and Hprt (sc-20975) (Santa Cruz).

    Techniques: Clonogenic Assay, Western Blot, Labeling, Radioactivity, Flow Cytometry, Staining, BrdU Incorporation Assay

    The S609A mutation inhibits activation of Src family kinases. ( a ) 1b9 or S609A cells were stimulated by ristocetin in the presence or absence of VWF, then solubilized and subjected to Western blot analysis with anti-Src and anti-phospho-Src family (pTyr416) antibody, respectively. The immunoblot is representative of 3 different experiments; ( b ) Quantitative data from 3 different experiments (mean ± SD) are demonstrated. The relative activation index of Src equals arbitrary quantitation of phospho-Src (pTyr416)/arbitrary quantitation of total Src.

    Journal: International Journal of Molecular Sciences

    Article Title: Role of 14-3-3ζ in Platelet Glycoprotein Ibα-von Willebrand Factor Interaction-Induced Signaling

    doi: 10.3390/ijms13055364

    Figure Lengend Snippet: The S609A mutation inhibits activation of Src family kinases. ( a ) 1b9 or S609A cells were stimulated by ristocetin in the presence or absence of VWF, then solubilized and subjected to Western blot analysis with anti-Src and anti-phospho-Src family (pTyr416) antibody, respectively. The immunoblot is representative of 3 different experiments; ( b ) Quantitative data from 3 different experiments (mean ± SD) are demonstrated. The relative activation index of Src equals arbitrary quantitation of phospho-Src (pTyr416)/arbitrary quantitation of total Src.

    Article Snippet: Anti-phospho-Src family (pTyr416) rabbit polyclonal antibody was from Cell Signaling Technology (Beverly, MA, USA).

    Techniques: Mutagenesis, Activation Assay, Western Blot, Quantitation Assay

    Animals were treated with 0.5% DMSO (control) or PD98059 at 300 µg/kg. Immunoblots and the respective densitometries are shown for p-ERK1/2, ERK1, p-Src and Src. Thirty µg of total protein were applied to each lane and each band is representative of one animal. β-actin was used as loading control. Relative densitometries as ratio of p-ERK/ERK1 and p-Src/Src are shown as fold change of control group and are presented by bars as means±SD (n = 3/group). *p<0.05 when comparing PD98059-treated group to control. Assays were conducted in duplicates.

    Journal: PLoS ONE

    Article Title: MAPK Signaling Pathway Regulates p27 Phosphorylation at Threonin 187 as Part of the Mechanism Triggered by Early-Weaning to Induce Cell Proliferation in Rat Gastric Mucosa

    doi: 10.1371/journal.pone.0066651

    Figure Lengend Snippet: Animals were treated with 0.5% DMSO (control) or PD98059 at 300 µg/kg. Immunoblots and the respective densitometries are shown for p-ERK1/2, ERK1, p-Src and Src. Thirty µg of total protein were applied to each lane and each band is representative of one animal. β-actin was used as loading control. Relative densitometries as ratio of p-ERK/ERK1 and p-Src/Src are shown as fold change of control group and are presented by bars as means±SD (n = 3/group). *p<0.05 when comparing PD98059-treated group to control. Assays were conducted in duplicates.

    Article Snippet: Samples were incubated overnight at 4°C with rabbit polyclonal antibodies to the signaling proteins ERK1 (1∶5000, sc-93, Santa Cruz Biotechnology) and phospho-Src (1∶300, #2101, Cell Signaling Technology), and to the cell cycle proteins cyclin E (1∶200, sc-481), CDK2 (1∶200, sc-163), cyclin D1 (1∶200, sc-718), CDK4 (1∶300, sc-601), p27 (1∶100, sc-528) (Santa Cruz Biotechnology), and phospho-p27 (T187, 1∶750, ab75908, ABCAM).

    Techniques: Western Blot